构建适合骨髓衍生肝干细胞移植的小鼠酒精性肝纤维化模型  

Construction of alcohol hepatic fibrosis models suitable for transplantation of bone marrow derived hepatic stem cells in mice

作　　者：苏俊[1] 廖彩仙[1] 周杰[1] 唐浩[1] 张守华[1] 张春兴[1] 

机构地区：[1]南方医科大学南方医院肝胆血管外科,广东省广州市510515

出　　处：《中国组织工程研究与临床康复》2007年第4期649-652,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金项目(30371385);广东省自然科学基金项目(04020419)~~

摘　　要：目的:建立适合骨髓衍生肝干细胞移植的小鼠酒精性肝纤维化模型,为移植骨髓衍生肝干细胞对酒精性肝损伤修复的研究提供进一步探讨的价值。方法:实验于2006-02/06在南方医科大学实验动物中心完成。取Balb/c雌性小鼠100只,SPF级,5～6周龄,体质量15～18g,随机区组法分为造模组92只,对照组8只。用白酒灌胃法建立小鼠酒精性肝纤维化模型。造模组于灌胃的第4,8,12,16周末分别随机选取6只小鼠,眼球采血后,颈椎脱臼处死,取出肝脏,以100g/L的甲醛固定,石蜡包埋切片,苏木精-伊红染色和苦味酸-酸性复红染色,光镜观察病理变化。血标本用于测定生化及肝纤维化指标。对照组以等量的蒸馏水灌胃,于实验的第16周末处死,处理同造模组。组间比较采用单因素方差分析。结果:纳入Balb/c雌性小鼠100只,实验中造模组共23只小鼠死亡,多为急性和亚急性死亡。对照组全部成活。①灌胃初期,造模组小鼠出现精神萎靡等现象,实验开始4周后上述症状减轻,对照组小鼠则食欲正常,无嗜睡现象。②在实验刚开始1周时小鼠体质量(15.67±0.98)g,与实验开始时(16.38±0.58)g相比呈明显负增长(P<0.05)。第4,8,12,16周称质量时,造模组小鼠体质量[(16.17±0.91),(17.23±1.39),(18.27±0.85),(19.31±0.95)g]均明显低于对照组[(18.31±0.57),(19.52±0.85),(20.56±0.86),(21.41±0.95)g],(P<0.05)。③对照组小鼠肝脏表面光滑细润、色红等。造模组小鼠肝脏色泽较正常肝脏暗淡,表面充血,肝脏呈增大趋势。造模组小鼠的肝重比(肝脏湿重/小鼠体质量×100%)均大于对照组。④造模组4周时可见轻度酒精性脂肪肝病理改变,12周时酒精性肝炎改变加重并出现了纤维组织增生,16周时肝纤维化已明显形成。⑤不同时间点与对照组相比,血清中天冬氨酸氨基转氨酶、丙氨酸氨基转氨酶、天冬氨酸氨基转氨酶/丙氨酸氨基转氨酶、三酰甘油均呈升高的AIM： To establish mouse alcohol hepatic fibrosis model for transplantation of bone marrow derived hepatic stem cells, so as to provide further approach for the research of repair of alcohol hepatic injury with bone marrow derived hepatic stem cells. METHODS： This experiment was conducted at the animal experiment center of Nanfang Medical University from February to June 2006. 100 Balb/c female mice of SPF level with 5-6 weeks old and 15-18 g were selected and randomly divided into model group （n =92） and control group （n =8）. The mice were intragastrically infused with distillate spirit to establish the model of alcohol hepatic fibrosis. Six mice in the model group were randomly selected and executed by damaging the cervical vertebra at the end of the 4^th, 8^th, 12^th and 16^th week of infusing spirit, respectively. Blood in the eyeball was drawn, and liver tissue was resected and fixed with 10% formalin, following by paraffin embedding for observation of pathological changes with light microscope after HE staining and trinitrophenol-acid fuchsin staining. The blood was used for measurement of biochemical and hepatic fibrosis indexes such as alanine aminotransferase （ALT）, aspartate aminotransferase （AST）, triglyceride （TG）, total protein, albumin （AL）, hyaluronic acid （HA） and laminin （LN）. The control group was given matching distilled water and sacrificed at the end of the 16^th week. Comparison between groups was performed by ANOVA. RESULTS： Among 100 Balb/c mice, 23 in the model group died commonly because of acute or subacute death. Mice in the control group were all alive. ①In the initial stage of intragastric infusion, mice in the model group appeared depressed and other symptoms, which were decreased after 4 weeks. Mice in the control group were eusitia and not drowsiness. The body mass weighed at the 1^st week （15.67±0.98） g was negatively grew compared with that at the beginning of the experiment （16.38±0.58 g, P 〈 0.05）. The body mass of the model g

关 键 词：肝疾病/酒精性 肝纤维化 病理学 动态观察 

分 类 号：R394.2[医药卫生—医学遗传学]